Total RNA was extracted from liver and ileum tissues using Eastep Super Total RNA Extraction Kit (Promega) and 1 μg of RNA was transcribed following the manufacturer's instruction by using the Reverse Transcription System Kit (Promega), which produced high‐quality complementary DNA. A real‐time PCR was performed on LightCyclerH 480 (Roche) using SYBR Green II Master (Takara). All mRNA quantification data were normalized to the housekeeping 36B4 gene. All samples were run in duplicate in a single 384‐well reaction plate, and data of the reaction were collected and analyzed according to the ∆∆CT method. Primers used in this study are listed in the Supplementary Materials.
Lightcyclerh 480
Manufactured by Roche
The LightCycler® 480 is a real-time PCR instrument designed for high-throughput and versatile quantitative and qualitative nucleic acid analysis. The instrument utilizes proprietary thermal block technology to provide accurate and reproducible results. It is compatible with a wide range of detection chemistries and can be used for various applications, including gene expression analysis, genotyping, and high-resolution melting analysis.
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2 protocols using lightcyclerh 480
1
Real-Time PCR Analysis of Gene Expression
2
RNA Extraction and Real-Time qPCR Analysis
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Total RNA was extracted from tissues using TRIzol Reagent (Invitrogen), and 1 μg of RNA was transcribed following the manufacturer's instruction by using the Reverse Transcription System Kit (Promega), which produced high-quality complementary DNA. A real-time polymerase chain reaction was performed on LightCyclerH 480 (Roche) using SYBR Green II Master (Takara). All mRNA quantification data were normalized to the housekeeping 36B4 gene. All samples were run in duplicate in a single 384-well reaction plate, and data of the reaction were collected and analyzed according to the ΔΔC T method. Primers used in this study are listed in Supplementary Table 1, see section on supplementary data given at the end of this article.
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